6-CHLORO-9-(1'-GLYCITYL) -1SOALLOXAZINES
Sept. 5, 1952
Methyl 3a-Acetoxy-l l-ketoetiocholanate (LXXXIV, R = R' = OMe). -Methyl 3,11-diketoetiocholanate (LXXXIII, R = H)I10 (40 mg.) dissolved in ethanol (6 cc.) a t 0' was reduced with sodium borohydride (40 mg.) in ethanol (1 cc.) a t 0' for 20 hours.111 Isolation with ether in the usual way gave a product which was dissolved in 90% methanol ( 5 cc.), digitonin (100 mg.) in 90% methanol ( 5 cc.) was added, and the cloudy solution was left for one hour. The solvent was evaporated a t the water pump, and the residue was well extracted with ether. Evaporation of the solvent gave a product (35 mg.) which when seeded with methyl 3cu-hydroxy-ll-ketoetiocholanate~1only partially solidified. It was dissolved in pyridine (1 cc.) and acetylated with acetic anhydride (0.5 cc.) in the usual way. The product was dissolved in benzene-petroleum ether (1 :1) and poured onto a short alumina column (ca. 1 g.), which was washed with the same solvent mixture (50 cc.) and then with methanol (30 cc.). The non-polar solvents gave a solid material (26 mg.) which on crystallization from etherpetroleum ether yielded methyl 3a-acetoxy-1 l-ketoetiocholanate (LXXXIV, R = Ac, R' = OMe) (18 mg.) as laths, m.p. 150-154" (Kof.). The methanol fraction yielded a crystalline material (12 mg.), m.p. 176-181' (Kof.), probably methyl 3 cy acetoxy - 11@ hydroxyetiocholanate"2 formed by reduction of both the keto groups of (LXXXIII, R = H). It was combined with the mother liquors from the crystallization of (LXXXIV, R = Ac, R ' = OMe), dissolved in acetic acid (0.5 cc.) and oxidized with 0.4 cc. of a 2% chromium trioxide-acetic acid solution a t 18' for 12 hours. Isolation with ether gave a solid residue which on
Ac,
-
-
(110) Lardon a n d Reichstein, footnote 61. A sample was kindly provided b y Dr. H. Heymann. (111) Cf.H e y m a n n a n d Fieser, THISJOURNAL, 7 3 , 5252 (1951). (112) Lardon and Reichstein, (footnote 109) give m.p. 183-185' for this substance.
[COSTRIAUTIOX FROM
THE
4251
crystallization from ether-petroleum ether gave a furthe: 9.5 mg. of (LXXXIV, R = Ac, R' = OMe), m.p. 148-153 (Kof.) (total vield 27.5 mz.. 61%). Further crvstallization raised the m.p. t o 15$-154.5"(Kof.). An authentic sample had m.p. 152.5-154.5" (Kof.),lIS and there was no depression on admixture.
We wish to express our very warm appreciation to our preparative assistants, Mrs. Dorothy Voitle and Mr. Irving Osvar. Their skill, hard work and enthusiasm were decisive factors for the successful outcome of this investigation. We are indebted to Dr. Ajay K. Bose and Dr. Richard B. Turner for valuable improvements in certain stages of the synthesis. For their very generous cooperation, we should like t o thank the Monsanto Chemical Company, who placed a t our disposal large quantities of 4methoxytoluquinone and the trans adduct (IX), and Merck and Company, Inc., who provided various crucial materials. Finally, we are grateful to Merck and Company, Inc., for their confidence in supporting our program a t its outset, and to Merck, Research Corporation, Eli Lilly and Company, and the United States Public Health Service for continued liberal financial support. (113) Inter d., v. Euw, Lardon a n d Reichstein (footnote 63) give m.p. 152-153'.
CAMBRIDGE, MASSACHUSETTS
RESEARCH LABORATORIES OF MERCK& Co., INC.]
Studies on Carcinolytic Compounds. IV.
6-Chloro-9-( 1'-glycity1)-isoalloxazines
BY CLIFFORD H. SHUNK, FRANK R. KONIUSZY AND KARLFOLKERS RECEIVED MARCH31, 1952 Eleven isoalloxazines in which the substituents in the 6-, 7- and 9-positions were varied have been prepared. Seven 6chloro-9-glycitylisoalloxazines,two 6-methyl-7-chloro-9-glycitylisoalloxazines and the 6-methyl and the 6-methoxy derivatives of 9-dulcitylisoalloxazine were synthesized by the reaction of alloxan with the diamine obtained by hydrogenation of the appropriately substituted 2-nitro-S-glycitylaniline. The isoalloxazines were tested for their effect in enhancing the rate of regression of lymphosarcoma (6C3H-ED) transplants in C3H mice maintained on a riboflavin deficient diet. 6-Chloro-9( 1'-D-sorbityl)-isoalloxazineappeared to show some activity in several tests. The other compounds showed questionable or negative results in single tests.
6,7-Dichloro-9-(1 '-D-sorbityl)-isoalloxazine' was D-mannity1)-aniline (111)) 4-chloro-2-nitro-N-( 1'found to be effective in enhancing the rate of re- L-arabity1)-aniline (IV), 4-chloro-2-nitro-N-( 1 ' - ~ gression of lymphosarcoma transplants in mice. arabity1)-aniline (V) , 4-chloro-2-nitro-N-( 1'-D-ribiAddition isoalloxazines in which the substituents in ty1)-aniline (VI) and 4-chloro-2-nitro-N- (1'-D-xylithe 6-, 7- and 9-positions are varied have been syn- ty1)-aniline (VII), respectively. Reaction of 2thesized. Seven 6-chloro-9-glycitylisoalloxazines,chloro-4-iodo-5-nitrotoluene, prepared by the diazotwo 6-methyl-7-chloro-9-glycitylisoalloxazines and tization of 4-amino-2-chloro-5-nitrotoluene5 folG-methyl- and 6-methoxy-9-dulcitylisoalloxazine lowed by treatment with potassium iodide, with Dwere prepared. glucamine and with D-galactamine yielded 3-chloro1-Chloro-4-iodo-3-nitrobenzene2 was heated in 4-methyl-B-nitro-N-(l'-~-sorbityl)-aniIine (VIII), pyridine with D-glucamine, D-galactamine,' D- and 3-chloro-4-methyl-6-nitro-N- (1'-D-dukityl) -anmannamine, L-arabinamine, D-arabhamine, D- iline (IX). 4-Methyl-2-nitro-N- (1'-D-dukityl) -anribamine3 and ~ - x y l a m i n egiving ~ 4-chloro-2-nitro- iline (X) and 4-methoxy-2-nitro-N-(l'-~-dulcityl) N- (l'-D-sorbityl)-aniline (I), 4-chloro-2-nitro-N- aniline (XI) were prepared by the reaction of D(1'-D-dulcityl)-aniline (11), 4-chloro-2-nitro-N-(1'- galactamine with 4-iodo-3-nitrotoluene6 and with (1) F. W. Holly, E. W. Peel, R. Mozingo a n d K. Folkers, THIS 4-iodo-3-nitroanisole,7 respectively. JOURNAL, 71, 5416 (1950). I n the condensation of the substituted iodoben(2) Korner, GQZZ.chim. i t d . , 4, 381 (1874). ( 3 ) F. W. Holly, E . W Peel, J. J. Cahill, F. R . Koniuzy and R. I'ulkers, THISJ O U R N A L , 1 4 , 4047 fl9.52). (4) F. W. IIolly, I
